Silver: Chemical compound screens: Difference between revisions
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*On the day of the screen, add 100 nl of library compounds to a final concentration of 10-40 µM. | *On the day of the screen, add 100 nl of library compounds to a final concentration of 10-40 µM. | ||
*Allow cells to incubate with compound for 1 hour at room temperature. | *Allow cells to incubate with compound for 1 hour at room temperature. | ||
*For visual-based immunostaining screens, wash cells with PBS and proceed through immunostaining procedure as described here. | *For visual-based immunostaining screens, wash cells with PBS and proceed through immunostaining procedure as described [[Silver: High-throughput immunostaining|here]]. |
Latest revision as of 14:10, 17 January 2006
Preparation
- Almost 24 hours prior to the screen, plate HeLa cells (or cells of choice)
- in 96- or 384-well Corning flat bottom plates (Corning 3614 or 3962, respectively)
- at a density of ~4000 cells/well in 50 µl of DMEM/10%FBS.
- If you have to infect cells to express your protein of interest, let infection proceed for ~24 hours prior to screen.
Compound Treatment
- On the day of the screen, add 100 nl of library compounds to a final concentration of 10-40 µM.
- Allow cells to incubate with compound for 1 hour at room temperature.
- For visual-based immunostaining screens, wash cells with PBS and proceed through immunostaining procedure as described here.